This kit provides a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating DNA as well as viral DNA from fresh, preserved or frozen urine samples from volumes ranging from 250 µL to 2 mL. All components for the purification are provided in one convenient and fast kit for the easy processing of small input volumes of bodily fluids.

The purified urine DNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive PCR and Southern Blot analysis, Microarrays and NGS.

Background

Recent evidence indicates that cell-free circulating DNA (cf-DNA) contains valuable information for the discovery of biomarkers that can help with early detection of certain cancer types and for monitoring the disease status. The advantage for using urine as a source for cancer biomarkers is that it can be acquired in large quantities without using invasive procedures. In addition, repeated sampling from the same individual is applicable, which facilitates longitudinal studies. There are many advantages favouring the use of urinary nucleic acids for cancer biomarker discovery over blood, tissue samples or other bodily fluids, including: (1) urine is non-infectious for HIV and less infectious for many other pathogens; (2) the profile of urinary nucleic acid is similar to that in plasma or serum; (3) nucleic acid purification from urine is technically much easier because of its low protein concentration (1000-fold lower than blood). Urine cell-free circulating DNA (cf-DNA) has been utilized for the early diagnosis, prognosis and monitoring of therapy for several cancer types and autoimmune diseases, as well as for investigating fetal DNA that is normally present in the maternal blood. This cf-DNA is usually present as short fragments, generally between 150 and 250 bp or its duplicates, and is derived either from the apoptotic process, necrotic process or from the fetus.

Kit Specifications

Minimum Urine Input

250 μL

Maximum Urine Input

2 mL

Size of DNA Purified

All sizes of
DNA ≥ 50 bp

Elution Volume

50-100 μL

Time to Complete 10 Purifications

15-20 minutes

Average Yields

Variable depending on specimen

*Please check page 6 of the product insert for average yields and the common DNA quantification methods.

Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature (15-25°C) for up to 2 years without showing any reduction in performance.

Norgen’s Urine Cell-Free Circulating DNA Purification Kits contain ready-to-use Proteinase K solution, which is dissolved in a specially prepared storage buffer. The Proteinase K is stable for up to 2 years after delivery when stored at room temperature. To prolong the lifetime of Proteinase K, storage at 2–8°C is recommended.